Increase in kidney Dh-S1P is not prevented by ABC294640 in lupus mice.

<p>After 10 weeks of ABC294640 or vehicle administration, kidneys were collected from MRL/MpJ or MLR/lpr mice. Kidney tissue was homogenized and analyzed for sphingolipid content by the Lipidomics Shared Resource at MUSC, <b>A</b>) sphingosine, <b>B</b>) S1P, <b>C</b>) Dh-sphingosine and <b>D</b>) Dh-S1P. Data represent mean ± SEM, n≥10; *p<0.05, **p<0.01, ***p<0.001 treated vs. MPJ.</p

Spleen weight and lymphocyte counts are not significantly altered by ABC294640.

<p>MRL/MpJ mice were treated with vehicle and MLR/lpr mice were treated with vehicle or ABC294640 for 10 weeks. <b>A</b>) Spleen weight was measured following euthanasia from mice at 20 weeks of age. <b>B</b>) Total B and <b>C</b>) T cell populations were analyzed using FACs flow cytometry. Data represent mean ± SEM, n≥10; **p<0.01, ***p<0.001 treated vs. MPJ.</p

ABC294640 does not prevent increases in urine thromboxane or albumin levels.

<p>After 10 weeks of ABC294640 or vehicle administration, 24-hour urine samples were collected from MRL/MpJ and MRL/lpr mice. <b>A</b>) Urine thromboxane and <b>B</b>) urine albumin levels were measured by ELISA. Data represent mean ± SEM, n≥10; *p<0.05 treated vs. MPJ.</p

ABC294640 prevents accumulation of Dh-S1P and S1P in circulation of lupus mice.

<p>After 10 weeks of ABC294640 or vehicle administration, whole blood was collected from MRL/MpJ and MRL/lpr mice. Blood was analyzed for sphingolipid content by the Lipidomics Shared Resource at MUSC, <b>A</b>) sphingosine, <b>B</b>) S1P, <b>C</b>) Dh-sphingosine and <b>D</b>) Dh-S1P. Data represent mean ±SEM, n≥10; *p<0.05, **p<0.01, ***p<0.001 treated vs. MPJ, unless otherwise indicated.</p

Glomerular pathology is not altered by treatment with ABC294640.

<p>MRL/MpJ mice were treated with vehicle and MLR/lpr mice were treated with vehicle or ABC294640 for 10 weeks. Kidneys were collected and sectioned for H&E staining. A blinded pathologist scored kidneys for <b>A</b>) vasculitis, interstitial <b>B</b>) focal or <b>C</b>) intensity, and glomerular <b>D</b>) focal or <b>D</b>) <b>i</b>ntensity scores. Data represent mean ± SEM, n≥10; *p<0.05, **p<0.01, ***p<0.001 treated vs. MPJ.</p

Lupus mice have elevated Dh-S1P levels in circulation.

<p>Serum was collected from MRL/lpr and MRL/MpJ mice and analyzed for sphingolipid content by the Lipidomics Shared Resource at MUSC, <b>A</b>) sphingosine, <b>B</b>) S1P, <b>C</b>) Dh-sphingosine and <b>D</b>) Dh-S1P. Data represent mean ± SEM, n = 6.</p

Sphingoid bases are elevated in kidney tissue from lupus mice.

<p>At 20 weeks of age, kidney tissue was collected from MRL/lpr and MRL/MpJ mice, homogenized and analyzed for sphingolipid content by the Lipidomics Shared Resource at MUSC, <b>A</b>) sphingosine, <b>B</b>) S1P, <b>C</b>) Dh-sphingosine and <b>D</b>) Dh-S1P. Data represent mean ± SEM, n = 6.</p

Mice with loss of SK1 in hematopoietic cells exhibit splenomegaly with DSS-induced colitis.

<p>Bone marrow transplants were performed with WT and SK1^−/− mice. Following 6 weeks of rest, mice were administered either regular drinking water or water containing 5% DSS for 5 days. <b>A</b>) Change in body weight was assessed. Data represent mean ±SD, n≥6 for each treatment group; (significance at Day 5 =  *WT^WTBM, **WT^SK1BM, **SK1^WTBM); *p<0.05, **p<0.01, ***p<0.001 vs strain Day 0. <b>B</b>) Spleen weight and <b>C</b>) colon length were assessed following treatment. <b>D</b>) Pathology damage scores were determined by a pathologist in a blinded fashion with H&E sections of colon tissue. Data represent mean ±SD, n≥6 for each treatment group; *p<0.05, **p<0.01, ***p<0.001 vs strain untreated, #p<0.05 as compared to WT^WTBM DSS treated, %p<0.05 as compared to SK1^SK1BM DSS. For the X-axis: regular text refers to the host genotype and the superscript to the bone marrow genotype.</p

Hematopoietic genotype determines circulating sphingolipid levels.

<p>Whole blood was collected and analyzed for sphingolipid content using ESI/MS/MS. <b>A and C</b>) Ceramide and <b>B and D</b>) S1P levels were normalized to volume. Data represent mean ±SD, n≥6 for each treatment group, *p<0.05, **p<0.01 as compared to both strains of mice with WT bone marrow. X-axis: regular text refers to the host genotype and the superscript to the bone marrow genotype.</p

p38 activation and IL-6 formation in a Gaucher’s disease mouse model.

<p>Proteins were extracted from tissues (brain, lung, and liver) of wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) and then subjected to immunoblot analysis with antibodies specific for phospho- p38 and β-actin. Equal amounts of protein were loaded in each lane, and the representative results of brain tissues are shown (<b>A</b>). Amounts of active/phospho-p38 were estimated by measuring the density of bands of phospho-p38 and expressed as arbitrary units (<b>B</b>). The data represent mean ± S.E. (n = 4–7). *, <i>p</i> < 0.02; **, <i>p</i> < 0.05. (<b>C</b>) mRNA was extracted from brain tissues from wild type and neuropathic Gaucher disease model V394L/PS-NA mice, and mRNAs of p38 isoforms and IL-6 were determined by the quantitative real time PCR. The data represent mean ± S.E. (n = 5). TNF-α, <i>p</i> < 0.0006 (*). (<b>D</b>) Serum IL-6 levels from wild type mice or Gaucher disease mouse models (V394L, D409H, and V394L/PS-NA) were determined by the ELISA system. The data represent mean ± S.E. Wild type, n = 21; V394L, n = 10; D409H, n = 7; V394L/PS-NA, n = 7. Wt <i>vs</i> V394L, <i>p</i> < 0.0006 (*); Wt <i>vs</i> D409, <i>p</i> < 0.03 (**); Wt <i>vs</i> V394L/PS-NA, <i>p</i> < 0.05 (***).</p